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1.
Anal Chem ; 88(7): 3826-35, 2016 Apr 05.
Artigo em Inglês | MEDLINE | ID: mdl-26959687

RESUMO

Here, we show results on X-ray absorption near edge structure spectroscopy in both transmission and X-ray fluorescence full-field mode (FF-XANES) at the calcium K-edge on human bone tissue in healthy and diseased conditions and for different tissue maturation stages. We observe that the dominating spectral differences originating from different tissue regions, which are well pronounced in the white line and postedge structures are associated with polarization effects. These polarization effects dominate the spectral variance and must be well understood and modeled before analyzing the very subtle spectral variations related to the bone tissue variations itself. However, these modulations in the fine structure of the spectra can potentially be of high interest to quantify orientations of the apatite crystals in highly structured tissue matrices such as bone. Due to the extremely short wavelengths of X-rays, FF-XANES overcomes the limited spatial resolution of other optical and spectroscopic techniques exploiting visible light. Since the field of view in FF-XANES is rather large the acquisition times for analyzing the same region are short compared to, for example, X-ray diffraction techniques. Our results on the angular absorption dependence were verified by both site-matched polarized Raman spectroscopy, which has been shown to be sensitive to the orientation of bone building blocks and by mathematical simulations of the angular absorbance dependence. As an outlook we further demonstrate the polarization based assessment of calcium-containing crystal orientation and specification of calcium in a beta-tricalcium phosphate (ß-Ca3(PO4)2 scaffold implanted into ovine bone. Regarding the use of XANES to assess chemical properties of Ca in human bone tissue our data suggest that neither the anatomical site (tibia vs jaw) nor pathology (healthy vs necrotic jaw bone tissue) affected the averaged spectral shape of the XANES spectra.


Assuntos
Cálcio/química , Osso Cortical/química , Minerais/química , Espectroscopia por Absorção de Raios X , Fosfatos de Cálcio/química , Simulação por Computador , Fluorescência , Humanos , Análise Espectral Raman , Raios X
2.
Part Fibre Toxicol ; 11: 13, 2014 Mar 25.
Artigo em Inglês | MEDLINE | ID: mdl-24666995

RESUMO

BACKGROUND: TiO2 particles are commonly used as dietary supplements and may contain up to 36% of nano-sized particles (TiO2-NPs). Still impact and translocation of NPs through the gut epithelium is poorly documented. RESULTS: We show that, in vivo and ex vivo, agglomerates of TiO2-NPs cross both the regular ileum epithelium and the follicle-associated epithelium (FAE) and alter the paracellular permeability of the ileum and colon epithelia. In vitro, they accumulate in M-cells and mucus-secreting cells, much less in enterocytes. They do not cause overt cytotoxicity or apoptosis. They translocate through a model of FAE only, but induce tight junctions remodeling in the regular ileum epithelium, which is a sign of integrity alteration and suggests paracellular passage of NPs. Finally we prove that TiO2-NPs do not dissolve when sequestered up to 24 h in gut cells. CONCLUSIONS: Taken together these data prove that TiO2-NPs would possibly translocate through both the regular epithelium lining the ileum and through Peyer's patches, would induce epithelium impairment, and would persist in gut cells where they would possibly induce chronic damage.


Assuntos
Epitélio/metabolismo , Trato Gastrointestinal/metabolismo , Nanopartículas/metabolismo , Nanopartículas/toxicidade , Titânio/farmacocinética , Titânio/toxicidade , Animais , Transporte Biológico , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Humanos , Absorção Intestinal , Camundongos , Microscopia Eletrônica de Transmissão , Nanopartículas/química , Reação em Cadeia da Polimerase em Tempo Real , Espectrometria por Raios X , Suspensões , Titânio/química , Espectroscopia por Absorção de Raios X
3.
Part Fibre Toxicol ; 11: 23, 2014 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-24885771

RESUMO

BACKGROUND: Although major concerns exist regarding the potential consequences of human exposure to nanoparticles (NP), no human toxicological data is currently available. To address this issue, we took welders, who present various adverse respiratory outcomes, as a model population of occupational exposure to NP.The aim of this study was to evaluate if welding fume-issued NP could be responsible, at least partially, in the lung alterations observed in welders. METHODS: A combination of imaging and material science techniques including ((scanning) transmission electron microscopy ((S)TEM), energy dispersive X-ray (EDX), and X-ray microfluorescence (µXRF)), was used to characterize NP content in lung tissue from 21 welders and 21 matched control patients. Representative NP were synthesized, and their effects on macrophage inflammatory secretome and migration were evaluated, together with the effect of this macrophage inflammatory secretome on human lung primary fibroblasts differentiation. RESULTS: Welding-related NP (Fe, Mn, Cr oxides essentially) were identified in lung tissue sections from welders, in macrophages present in the alveolar lumen and in fibrous regions. In vitro macrophage exposure to representative NP (Fe2O3, Fe3O4, MnFe2O4 and CrOOH) induced the production of a pro-inflammatory secretome (increased production of CXCL-8, IL-1ß, TNF-α, CCL-2, -3, -4, and to a lesser extent IL-6, CCL-7 and -22), and all but Fe3O4 NP induce an increased migration of macrophages (Boyden chamber). There was no effect of NP-exposed macrophage secretome on human primary lung fibroblasts differentiation. CONCLUSIONS: Altogether, the data reported here strongly suggest that welding-related NP could be responsible, at least in part, for the pulmonary inflammation observed in welders. These results provide therefore the first evidence of a link between human exposure to NP and long-term pulmonary effects.


Assuntos
Pulmão/patologia , Nanopartículas Metálicas/toxicidade , Doenças Profissionais/patologia , Óxidos/toxicidade , Soldagem , Idoso , Movimento Celular/efeitos dos fármacos , Citocinas/metabolismo , Feminino , Fibroblastos/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Exposição por Inalação , Pulmão/metabolismo , Macrófagos/efeitos dos fármacos , Masculino , Pessoa de Meia-Idade , Exposição Ocupacional , Fumar/efeitos adversos , Fumar/patologia , Fixação de Tecidos
4.
Part Fibre Toxicol ; 10: 24, 2013 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-23800198

RESUMO

BACKGROUND: Carbon nanotubes (CNT) are a family of materials featuring a large range of length, diameter, numbers of walls and, quite often metallic impurities coming from the catalyst used for their synthesis. They exhibit unique physical properties, which have already led to an extensive development of CNT for numerous applications. Because of this development and the resulting potential increase of human exposure, an important body of literature has been published with the aim to evaluate the health impact of CNT. However, despite evidences of uptake and long-term persistence of CNT within macrophages and the central role of those cells in the CNT-induced pulmonary inflammatory response, a limited amount of data is available so far on the CNT fate inside macrophages. Therefore, the overall aim of our study was to investigate the fate of pristine single walled CNT (SWCNT) after their internalization by macrophages. METHODS: To achieve our aim, we used a broad range of techniques that aimed at getting a comprehensive characterization of the SWCNT and their catalyst residues before and after exposure of murine macrophages: X-ray diffraction (XRD), High Resolution (HR) Transmission Electron Microscopy (TEM), High Angle Annular Dark Field-Scanning TEM (HAADF-STEM) coupled to Electron Energy Loss Spectroscopy (EELS), as well as micro-X-ray fluorescence mapping (µXRF), using synchrotron radiation. RESULTS: We showed 1) the rapid detachment of part of the iron nanoparticles initially attached to SWCNT which appeared as free iron nanoparticles in the cytoplasm and nucleus of CNT-exposed murine macrophages, and 2) that blockade of intracellular lysosomal acidification prevented iron nanoparticles detachment from CNT bundles and protected cells from CNT downstream toxicity. CONCLUSIONS: The present results, while obtained with pristine SWCNT, could likely be extended to other catalyst-containing nanomaterials and surely open new ways in the interpretation and understanding of CNT toxicity.


Assuntos
Compostos de Ferro/metabolismo , Macrófagos/metabolismo , Nanopartículas Metálicas , Nanotubos de Carbono/análise , Animais , Catepsina B/metabolismo , Linhagem Celular , Concentração de Íons de Hidrogênio , Compostos de Ferro/toxicidade , Proteínas de Membrana Lisossomal/metabolismo , Lisossomos/metabolismo , Macrolídeos/farmacologia , Macrófagos/efeitos dos fármacos , Camundongos , Microscopia Eletrônica de Transmissão , Nanotubos de Carbono/toxicidade , Espectrometria por Raios X , Espectroscopia de Perda de Energia de Elétrons , Síncrotrons , Difração de Raios X
5.
Biomed Opt Express ; 13(3): 1620-1639, 2022 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-35415001

RESUMO

White-matter injury leads to severe functional loss in many neurological diseases. Myelin staining on histological samples is the most common technique to investigate white-matter fibers. However, tissue processing and sectioning may affect the reliability of 3D volumetric assessments. The purpose of this study was to propose an approach that enables myelin fibers to be mapped in the whole rodent brain with microscopic resolution and without the need for strenuous staining. With this aim, we coupled in-line (propagation-based) X-ray phase-contrast tomography (XPCT) to ethanol-induced brain sample dehydration. We here provide the proof-of-concept that this approach enhances myelinated axons in rodent and human brain tissue. In addition, we demonstrated that white-matter injuries could be detected and quantified with this approach, using three animal models: ischemic stroke, premature birth and multiple sclerosis. Furthermore, in analogy to diffusion tensor imaging (DTI), we retrieved fiber directions and DTI-like diffusion metrics from our XPCT data to quantitatively characterize white-matter microstructure. Finally, we showed that this non-destructive approach was compatible with subsequent complementary brain sample analysis by conventional histology. In-line XPCT might thus become a novel gold-standard for investigating white-matter injury in the intact brain. This is Part I of a series of two articles reporting the value of in-line XPCT for virtual histology of the brain; Part II shows how in-line XPCT enables the whole-brain 3D morphometric analysis of amyloid- ß (A ß ) plaques.

6.
Biophys J ; 101(2): 486-93, 2011 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-21767502

RESUMO

Amyloidoses are increasingly recognized as a major public health concern in Western countries. All amyloidoses share common morphological, structural, and tinctorial properties. These consist of staining by specific dyes, a fibrillar aspect in electron microscopy and a typical cross-ß folding in x-ray diffraction patterns. Most studies that aim at deciphering the amyloid structure rely on fibers generated in vitro or extracted from tissues using protocols that may modify their intrinsic structure. Therefore, the fine details of the in situ architecture of the deposits remain unknown. Here, we present to our knowledge the first data obtained on ex vivo human renal tissue sections using x-ray microdiffraction. The typical cross-ß features from fixed paraffin-embedded samples are similar to those formed in vitro or extracted from tissues. Moreover, the fiber orientation maps obtained across glomerular sections reveal an intrinsic texture that is correlated with the glomerulus morphology. These results are of the highest importance to understanding the formation of amyloid deposits and are thus expected to trigger new incentives for tissue investigation. Moreover, the access to intrinsic structural parameters such as fiber size and orientation using synchrotron x-ray microdiffraction, could provide valuable information concerning in situ mechanisms and deposit formation with potential benefits for diagnostic and therapeutic purposes.


Assuntos
Amiloide/química , Síncrotrons , Difração de Raios X , Amiloidose/metabolismo , Amiloidose/patologia , Anisotropia , Humanos , Glomérulos Renais/metabolismo , Glomérulos Renais/patologia
7.
Phys Med Biol ; 66(6): 065005, 2021 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-32268312

RESUMO

X-ray phase contrast imaging can provide improved or complementary information to traditional attenuation-based X-ray imaging, making the field a vast and rapidly evolving research subject. X-ray speckle-based imaging (SBI) is one phase-contrast imaging approach that has shown significant potential in providing both high sensitivity and high resolution while using a very simple experimental setup. With the aim of transferring such phase-contrast-based imaging techniques from synchrotron to laboratory X-ray sources, the issue of the deposited radiation dose still remains to be addressed. In this work, we experimentally and quantitatively compare the results from three different SBI phase retrieval algorithms using both phantoms and biological samples in order to infer the optimal configuration. The results obtained using a synchrotron beam suggest that the technique based on optical flow conservation achieves the most accurate retrieval from the lowest number of sample exposures. This constitutes an important step toward the possibility of transferring SBI into the clinic.


Assuntos
Microscopia de Contraste de Fase/métodos , Doses de Radiação , Radiografia/métodos , Radiometria/métodos , Síncrotrons , Raios X , Algoritmos , Desenho de Equipamento , Humanos , Variações Dependentes do Observador , Imagens de Fantasmas , Controle de Qualidade
8.
Sci Rep ; 10(1): 1911, 2020 02 05.
Artigo em Inglês | MEDLINE | ID: mdl-32024864

RESUMO

X-ray Phase Contrast Imaging (PCI) is an emerging modality whose availability in clinics for mammography and lung imaging is expected to materialize within the coming years. In this study, we evaluate the PCI Computed Tomography (PCI-CT) performances with respect to current conventional imaging modalities in the context of osteo-articular disorders diagnosis. X-ray PCI-CT was performed on 3 cadaveric human hands and wrists using a synchrotron beam. Conventional CT, MRI and Ultrasound were also performed on these three samples using routine procedures as well as research protocols. Six radiologists and rheumatologists independently evaluated qualitatively and semi quantitatively the 3D images' quality. Medical interpretations were also made from the images. PCI-CT allows the simultaneous visualization of both the high absorbing and the softer tissues. The 6 reader evaluations characterized PCI-CT as a visualization tool with improved performances for all tissue types (significant p-values), which provides sharper outlines and clearer internal structures than images obtained using conventional modalities. The PCI-CT images contain overall more information, especially at smaller scales with for instance more visible micro-calcifications in our chondrocalcinosis case. Despite a reduced number of samples used, this pilot study highlights the possible medical benefits of PCI for osteo-articular disorders evaluation. Although PCI-CT is not yet available in hospitals, the improved visualization capabilities demonstrated so far and the enhanced tissue measurement quality let suggest strong diagnosis benefits for rheumatology in case of a widespread application of PCI.


Assuntos
Condrocalcinose/diagnóstico , Articulação da Mão/diagnóstico por imagem , Osteoartrite/diagnóstico , Tomografia Computadorizada por Raios X/métodos , Cadáver , Humanos , Imageamento por Ressonância Magnética , Projetos Piloto , Síncrotrons , Tomografia Computadorizada por Raios X/instrumentação
9.
Sci Rep ; 8(1): 14003, 2018 09 18.
Artigo em Inglês | MEDLINE | ID: mdl-30228304

RESUMO

Human vocal folds possess outstanding abilities to endure large, reversible deformations and to vibrate up to more than thousand cycles per second. This unique performance mainly results from their complex specific 3D and multiscale structure, which is very difficult to investigate experimentally and still presents challenges using either confocal microscopy, MRI or X-ray microtomography in absorption mode. To circumvent these difficulties, we used high-resolution synchrotron X-ray microtomography with phase retrieval and report the first ex vivo 3D images of human vocal-fold tissues at multiple scales. Various relevant descriptors of structure were extracted from the images: geometry of vocal folds at rest or in a stretched phonatory-like position, shape and size of their layered fibrous architectures, orientation, shape and size of the muscle fibres as well as the set of collagen and elastin fibre bundles constituting these layers. The developed methodology opens a promising insight into voice biomechanics, which will allow further assessment of the micromechanics of the vocal folds and their vibratory properties. This will then provide valuable guidelines for the design of new mimetic biomaterials for the next generation of artificial larynges.


Assuntos
Imageamento Tridimensional/métodos , Síncrotrons/instrumentação , Prega Vocal/anatomia & histologia , Prega Vocal/fisiologia , Microtomografia por Raio-X/métodos , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Modelos Anatômicos , Fonação , Voz
10.
J Mol Biol ; 356(4): 843-9, 2006 Mar 03.
Artigo em Inglês | MEDLINE | ID: mdl-16405906

RESUMO

The soluble protein Ure2p from the yeast Saccharomyces cerevisiae assembles in vitro into straight and insoluble protein fibrils, through subtle changes of conformation. Whereas the structure of soluble Ure2p has been revealed by X-ray crystallography, further characterization of the structure of insoluble Ure2p fibrils is needed. We performed X-ray absorption near-edge spectroscopy (XANES) at the sulfur K-edge to probe the state of Cys221 in the fibrillar form of Ure2pC221 and provide structural information on the structure of Ure2p within fibrils. Although the Ure2p dimer dissociation into its constituent monomers has proven to be a prerequisite for assembly into fibrils, we showed the ability of every Ure2pC221 monomer to establish disulfide bonds upon incubation of the fibrils under oxidizing conditions. Our result indicates either that the constituent unit of the fibrillar form of the protein is a dimeric Ure2p or that the fibrils are made of protofilaments assembled in such a way that the residue C221 from a Ure2p molecule in one protofilament is located in the vicinity of a C221 residue from another molecule belonging to a neighbor protofilament.


Assuntos
Príons/química , Conformação Proteica , Proteínas de Saccharomyces cerevisiae/química , Enxofre/química , Dissulfetos/química , Glutationa Peroxidase , Oxirredução , Príons/genética , Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/genética , Análise Espectral/métodos
11.
Zoolog Sci ; 19(1): 27-35, 2002 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-12025401

RESUMO

Some ascidians (sea squirts) accumulate the transitional metal vanadium in their blood cells at concentrations of up to 350 mM, about 10(7) times its concentration found in seawater. There are approximately 10 different types of blood cell in ascidians. The identity of the true vanadium-containing blood cell (vanadocyte) is controversial and little is known about the subcellular distribution of vanadium. A scanning x-ray microscope installed at the ID21 beamline of the European Synchroton Radiation Facility to visualize vanadium in ascidian blood cells. Without fixation, freezing or staining realized the visualization of vanadium localized in living signet ring cells and vacuolated amoebocytes of two vanadium-rich ascidian species, Phallusia mammillata and Ascidia sydneiensis samea. A combination of transmission and fluorescence images of signet ring cells suggested that in both species the vacuoles contain vanadium.


Assuntos
Células Sanguíneas/química , Células Sanguíneas/ultraestrutura , Microscopia/métodos , Urocordados/citologia , Urocordados/metabolismo , Vanádio/sangue , Animais , Sobrevivência Celular , Liofilização , Líquido Intracelular/metabolismo , Microscopia/instrumentação , Síncrotrons , Urocordados/classificação , Vacúolos/metabolismo , Raios X
12.
Nanotoxicology ; 8 Suppl 1: 167-76, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24359331

RESUMO

We assessed the biodistribution and in situ speciation of sub-lethal concentrations of citrate-coated silver nanoparticles and dissolved silver within Fundulus heteroclitus embryos. Using a thorough physico-chemical characterization, we studied the role of salinity on both uptake and in situ speciation. The Ag uptake or adsorption on the chorion was reduced by 2.3-fold for Ag NPs, and 2.9-fold for AgNO3 in estuarine water (10‰ ASW) compared to deionized water (0‰ ASW). Between 58% and 85% of the silver was localized on/in the chorion and formed patches between 20 and 80 µm. More than a physical barrier, the chorion was found to be a chemically reactive membrane controlling the in situ speciation of silver. A strong complexation of the Cit-Ag NPs with the thiolated groups of proteins or enzymes of the chorion was responsible for the oxidation of 48 ± 5% of the Ag(0) into Ag((I))-S species at 0‰ ASW. However, at 10‰ ASW, the presence of Cl(-) ions at the surface of Ag NPs slow down this oxidation. For the dissolved silver, we observed that in deionized water 69 ± 7% of Ag(+) taken up by the chorion was complexed by the thiolated molecules while the others 30 ± 3% were reduced into Ag(0) likely via interaction with the hemiacetal-reducing ends of polysaccharides of the chorion.


Assuntos
Fundulidae/metabolismo , Nanopartículas/metabolismo , Prata/química , Cloreto de Sódio/metabolismo , Animais , Fundulidae/embriologia , Nanopartículas/química
13.
Int J Pharm ; 453(2): 448-53, 2013 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-23792043

RESUMO

Curing is generally required to stabilize film coating from aqueous polymer dispersion. This post-coating drying step is traditionally carried out in static conditions, requiring the transfer of solid dosage forms to an oven. But, curing operation performed directly inside the coating equipment stands for an attractive industrial application. Recently, the use of various advanced physico-chemical characterization techniques i.e., X-ray micro-computed tomography, vibrational spectroscopies (near infrared and Raman) and X-ray microdiffraction, allowed new insights into the film-coating structures of dynamically cured tablets. Dynamic curing end-point was efficiently determined after 4h. The aim of the present work was to elucidate the influence of curing conditions on film-coating structures. Results demonstrated that 24h of static curing and 4h of dynamic curing, both performed at 60°C and ambient relative humidity, led to similar coating layers in terms of drug release properties, porosity, water content, structural rearrangement of polymer chains and crystalline distribution. Furthermore, X-ray microdiffraction measurements pointed out different crystalline coating compositions depending on sample storage time. An aging mechanism might have occur during storage, resulting in the crystallization and the upward migration of cetyl alcohol, coupled to the downward migration of crystalline sodium lauryl sulfate within the coating layer. Interestingly, this new study clearly provided further knowledge into film-coating structures after a curing step and confirmed that curing operation could be performed in dynamic conditions.


Assuntos
Composição de Medicamentos/métodos , Comprimidos/química , Cristalização , Álcoois Graxos/química , Temperatura Alta , Dodecilsulfato de Sódio/química , Espectroscopia de Luz Próxima ao Infravermelho , Análise Espectral Raman , Difração de Raios X , Microtomografia por Raio-X
14.
Sci Total Environ ; 431: 197-208, 2012 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-22684121

RESUMO

Intensive production of TiO(2) nanoparticles (TiO(2)-NPs) would lead to their release in the environment. Their ecotoxicological impact is still poorly documented, while their use in commercial goods is constantly increasing. In this study we compare root accumulation and root-to-shoot translocation in wheat of anatase and rutile TiO(2)-NPs with diameters ranging from 14 nm to 655 nm, prepared in water. NP distribution in plant tissues was mapped by synchrotron-radiation micro-X-ray fluorescence, observed by transmission electron microscopy and quantified in the different compartments of plant roots by micro-particle-induced X-ray emission. Our results provide evidence that the smallest TiO(2)-NPs accumulate in roots and distribute through whole plant tissues without dissolution or crystal phase modification. We suggest a threshold diameter, 140 nm, above which NPs are no longer accumulated in wheat roots, as well as a threshold diameter, 36 nm, above which NPs are accumulated in wheat root parenchyma but do not reach the stele and consequently do not translocate to the shoot. This accumulation does not impact wheat seed germination, biomass and transpiration. It does not induce any modification of photosynthesis nor induce oxidative stress. However exposure of wheat plantlets to the smallest NPs during the first stages of development causes an increase of root elongation. Collectively, these data suggest that only the smallest TiO(2)-NPs may be accumulated in wheat plants, although in limited amounts and that their impact is moderate.


Assuntos
Poluentes Ambientais/farmacocinética , Nanopartículas , Titânio/farmacocinética , Triticum/metabolismo , Ecotoxicologia/métodos , Germinação , Microscopia Eletrônica de Transmissão , Tamanho da Partícula , Folhas de Planta/metabolismo , Raízes de Plantas/metabolismo , Brotos de Planta/metabolismo , Espectrometria por Raios X/métodos , Distribuição Tecidual , Triticum/crescimento & desenvolvimento
15.
Chem Res Toxicol ; 18(10): 1512-9, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16533014

RESUMO

Hexavalent chromium compounds are known carcinogens for the respiratory tract in humans. The mechanism of cell transformation by hexavalent chromium compounds is not fully understood although a role for intracellular reduction is sought. The aim of this study was to determine the distribution of Cr valence states in human cells after in vitro exposure to soluble or particulate chromium compounds. A synchrotron X-ray-based microprobe was used to investigate the cellular reduction of Cr(VI) and to image chromium oxidation states in cells. It was shown that soluble Cr(VI) compounds are fully reduced to Cr(III) in cells. Cr(III) is homogeneously distributed within the cell volume and therefore present within the nucleus. In the case of low solubility particulate chromate compounds, Cr(VI) can coexist in the cell environment, as particles in the perinuclear region, together with intracellular and intranuclear Cr(III). Chemical distribution maps also suggest that intracellular Cr(III) originates from extracellular dissolution and reduction of lead chromate rather than from intracellular engulfed particles. The possible stronger carcinogenicity of low solubility chromate vs soluble chromate compounds may derive from the combinative genotoxic effects of intranuclear Cr(III) and the persistent exposure to a strong oxidant, Cr(VI).


Assuntos
Carcinógenos/farmacologia , Cromatos/farmacologia , Microanálise por Sonda Eletrônica , Animais , Células CHO , Carcinógenos/toxicidade , Cromatos/toxicidade , Cromo/química , Cromo/metabolismo , Cricetinae , Cricetulus , Chumbo/farmacologia , Oxirredução , Compostos de Sódio/farmacologia , Solubilidade
16.
J Biol Chem ; 278(13): 11654-60, 2003 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-12538647

RESUMO

Transthyretin (TTR) amyloid fibrils are the main component of the amyloid deposits occurring in Familial Amyloidotic Polyneuropathy patients. This is 1 of 20 human proteins leading to protein aggregation disorders such as Alzheimer's and Creutzfeldt-Jakob diseases. The structural details concerning the association of the protein molecules are essential for a better understanding of the disease and consequently the design of new strategies for diagnosis and therapeutics. Disulfide bonds are frequently considered essential for the stability of protein aggregates and since in the TTR monomers there is one cysteine residue, it is important to determine unambiguously the redox state of sulfur present in the fibrils. In this work we used x-ray spectroscopy to further characterize TTR amyloid fibrils. The sulfur K-edge absorption spectra for the wild type and some amyloidogenic TTR variants in the soluble and fibrillar forms were analyzed. Whereas in the soluble proteins the thiol group from cysteine (R-SH) and the thioether group from methionine (R-S-CH(3)) are the most abundant forms, in the TTR fibrils there is a significant oxidation of sulfur to the sulfonate form in the cysteine residue and a partial oxidation of sulfur to sulfoxide in the methionine residues. Further interpretation of the data reveals that there are no disulfide bridges in the fibrillar samples and suggest conformational changes in the TTR molecule, namely in strand A and/or in its vicinity, upon fibril formation.


Assuntos
Pré-Albumina/metabolismo , Enxofre/metabolismo , Benzotiazóis , Microanálise por Sonda Eletrônica , Modelos Moleculares , Oxirredução , Pré-Albumina/química , Ligação Proteica , Conformação Proteica , Tiazóis/metabolismo , Difração de Raios X
17.
J Opt Soc Am A Opt Image Sci Vis ; 19(4): 797-806, 2002 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-11934173

RESUMO

X-ray imaging in differential interference contrast (DIC) with submicrometer optical resolution was performed by using a twin zone plate (TZP) setup generating focal spots closely spaced within the TZP spatial resolution of 160 nm. Optical path differences introduced by the sample are recorded by a CCD camera in a standard full-field imaging and by an aperture photodiode in a standard scanning transmission x-ray microscope. Applying this x-ray DIC technique, we demonstrate for both the full-field imaging and scanning x-ray microscope methods a drastic increase in image contrast (approximately 20x) for a low-absorbing specimen, similar to the Nomarski DIC method for visible-light microscopy.

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